Figure 4.

CFTR function in primary differentiated NHBE:CFBE cell mixed culture and CFBE cell models

(A and B) Example I_sc traces from NHBE:CFBE cells and (A) from transduced CFBE cells (B; percent final transduction for individual CFTR variants are given) after incubation with CFS and addition of specific activators and inhibitors of ion transport (as below). (C and D) ΔI_sc plotted against percent final NHBE after addition of (C) CFTR_inh172 (10 μM) and summarized in (D) as ΔI_sc per percent final NHBE or percent transduced cells in the culture (ΔI_sc/% final). The dotted horizontal lines in (C) mark ASL height in 100% NHBE cells in the presence of normal lung sputum (healthy) or presence of CFS. (E) TEER/percent final transduced for WT-CFTR and hCAI CFTR. Treatments were compared by unpaired t test. ∗p < 0.05 (n = 8–12 from 3 CF donors). (F) I_sc traces from CFBE cells transduced with WT or hCAI CFTRs or GFP alone before and after addition of a basolateral-to-apical Cl⁻ gradient and specific activators and inhibitors of ion transport: amiloride (10 μM), FSK (10 μM), CFTR_inh172 (10 μM), and UTP (10 μM). All drugs were added apically with exception of FSK, which was added bilaterally. Direction of chloride concentration gradients (using physiological Cl⁻ and Cl⁻-free buffers) are indicated, and the duration of application is presented as gray bars. ΔI_sc/percent final transduced in response to FSK and CFTR_inh172 is given in a table below the graph.